Therapeutic strategies for prevention and treatment of alzheimer's disease
a technology for alzheimer's disease and therapeutic strategies, applied in the field of new drugs, can solve the problems that the accumulation of alzheimer's protein does not have any effect on the cognitive status or functional capacity of the brain, and the disease (ad) is becoming a major health problem, and achieves the effect of being less effectiv
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Procedures Used to Test for the Presence of a Cell Cycle Regulatory Defect at the G1 / S Transition
[0103]The following methods of testing for the presence of a cell cycle regulatory defect at the G1 / S transition may be used “diagnostically” in order to test whether a given cell exhibits the regulatory defect and may also form the basis of compound screening methods. For compound screening applications, candidate compounds may be added before the cells are treated to induce cell division and elicit cell cycle arrest.
[0104]Method (A): (i) Induce the cells to divide (e.g., by the addition of a mitogenic stimulus, for example one or more growth factors), then (ii) elicit cell cycle arrest by addition of a cell division inhibitor substance (most preferably a specific G1 inhibitor, for example rapamycin), and (iii) test the responsiveness of the cells' G1 / S cell cycle regulatory mechanisms to the addition of the cell division inhibitor substance, in the presence or the absence of a candidat...
example 2
Protocols for Diagnostic Tests
[0146]The following are specific protocols for separation and culture of lymphocytes, induction of cell division, induction of cell cycle arrest by either treatment with a cell division inhibitor or H2O2-induced hypoxia, BrdU incorporation / FACS analysis and MTT survival assay. These protocols, or minor adaptations thereof, may all be used diagnostically to test for the presence of a regulatory defect at the G1 / S transition and in compound screening assays.
Lymphocyte Separation
[0147]Blood is collected in lithium heparin or EDTA vacutainers. Lymphocytes are isolated according to a standard protocol using Ficoll (Sigma). In order to standardize the culture methods for all patients, the separated lymphocytes are frozen and stored for further analysis.
[0148]When lymphocytes are needed for culture, they are thawed in a 37° C. water bath and washed twice in RPMI (any medium or buffer which supports lymphocyte survival may be used to wash the cells with equival...
example 3
Examining the Effects of Cell Cycle Inhibitor Drugs on Cell Division Kinetics and Alzheimer's-Type Protein Expression in the SH-SY5Y Neuroblastoma Cell Line
[0155]The object of this study was to demonstrate that exemplary cell cycle inhibitor drugs are able to reduce indicators of Alzheimer's disease pathology in an in vitro model of Alzheimer's disease. There is currently no available animal model for sporadic Alzheimer's disease. However, the SH-SY5Y neuroblastoma cell line (ATCC accession number CRL-2266) is accepted as a suitable in vitro model.
[0156]SH-SY5Y cells were treated with three candidate agents: rapamycin (Rapa) at 100 ng / ml, doxorubicin (Doxo) at 1 μM and Dexrazoxane (DexRaz) at 200 μM and then tested one day (d1) and two days (d2) after treatment for the following 7 parameters, which are indicative of cell cycle defects and “AD-type” pathology:
Parameters Indicative of Changes in Cell Cycle / Cell Division Kinetics:
[0157]1) Cell survival and proliferation[0158]2) Apoptos...
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